Target Sp15 - Acetyl-CoA carboxylase (Clostridium botulinum)

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Format for Individual Target pages (copy this list to new Target page and then fill in for your target):

*Target (protein/gene name): Acetyl-CoA carboxylase
*NCBI Gene # or RefSeq#: 441770
*Protein ID (NP or XP #) or Wolbachia#:
*Organism (including strain): Clostridium botulinum (strain ATCC 19397/Type A)
Etiologic Risk Group (see link below):
*/Disease Information (sort of like the Intro to your Mini Research Write up):
Clostridium botulinum is a gram-positive, rod-shaped bacteria responsible for botulism. The toxin caused by C. botulinum affects nerves and, if untreated, can cause paralysis and respiratory failure. Transmission is usually foodborne and can come from home-canned foods with low acid content or sealed containers.
Link to TDR Targets page (if present):
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) http://www.uniprot.org/uniprot/A7FPN8
Essentiality of this protein:
Enzyme is essential in fatty acid and phospholipid metabolism in C. botulinum. Essentiality was determined for C. botulinum and the homology between the selected protein and human orthologs was taken into account. The enzyme is a non-human homologous sequence.
http://www.sciencedirect.com/science/article/pii/S0888754314000664 (paper determining the essentiality of different proteins in C. botulinum)
Is it a monomer or multimer as biological unit? (make prediction at http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html): multimer
Complex of proteins?: complex
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
Inhibition of acetyl-CoA carboxylase (ACC) by isozyme-specific antisense oligonucleotides or isozyme-nonselective ACC inhibitors.
http://www.ncbi.nlm.nih.gov/pubmed/18221116
Inhibition of acetyl-CoA carboxylase is also inhibited by nanomolar concentrations of both haloxyfop and tralkoxydim.
http://www.plantphysiol.org/content/86/1/10.full.pdf

*EC#: 6.4.1.2
Link to BRENDA EC# page:
http://www.brenda-enzymes.info/enzyme.php?ecno=6.4.1.2
reaction.png

Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
Spectrophotometric assay coupling the carboxylation of acetyl-CoA to NADPH-dependent reduction of malonyl-CoA.
http://www.ncbi.nlm.nih.gov/pubmed/21138728

Structure (PDB or Homology model)
PDB structure:
http://www.rcsb.org/pdb/explore/explore.do?structureId=1OD4

Current Inhibitors: 589
Expression Information (has it been expressed in bacterial cells): has been expressed in E. Coli
http://www.ncbi.nlm.nih.gov/pubmed/14594796
Purification Method: Myc-5 IgG affinity column
http://www.sciencedirect.com/science/article/pii/S1046592806001781
Image of protein (PyMol with features delineated and shown separately):
acetyl coa.png
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): 
MDFKGIENLIKAMSESNLSSMDIEYNGIAIKMKKENNKIYKQETISQEYEKENRDNIVEE
KKLDLLSNEEKTGIAIADNLIEIVSPIVGTFYESPGVDKKPYAKAGDKVKKGDTVCIVEA
MKVMNEIEAEVDGEIVEVLVENEQMVQYGEVLFKIKPL
*length of your protein in Amino Acids: 158
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website:
17.85449 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 8940
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
Graph.gif
*CDS Gene Sequence (paste as text only):
gi 261417741 4 MKIQEIRELIRLVDQSSIDEFVYEQGETKVHMKK.[16].VAPP.[9].PAPAAPP.[4].ASEAPAPKA 86 gi 146319455 1 MNITEIKDLMSQFDQSSLREFSYSNAGEILHFSK.[12].EAPL.[9].PVPAVEMSEPAPAEAG 75 gi 146321652 1 MNITEIKDLMSQFDQSSLREFSYSNAGEILHFSK.[14].PLAP.[7].PVPAVEMSEPAPAEAG 75 gi 253752470 1 MNITEIKDLMSQFDQSSLREFSYSNAGEILHFSK.[14].PLAP.[7].PVPAVEMSEPAPAEAG 75 gi 253754296 1 MNITEIKDLMSQFDQSSLREFSYSNAGEILHFSK.[14].PLAP.[7].PVPAVEMSEPAPAEAG 75 gi 253756230 1 MNITEIKDLMSQFDQSSLREFSYSNAGEILHFSK.[14].PLAP.[7].PVPAVEMSEPAPAEAG 75 gi 288904558 2 VNISEVKDLMAQFDQSSLREFSFKTGEAELTFSK.[17].AAPVEVAATAPVAPA.[1].AAS.[1].SE 74 gi 55820480 1 MNISEIKDLLAQFDTSTLREFSYKNNGEELNLSK.[22].QAPV.[1].PVAAPAA.[1].ETPA.[1].PVEEA 79 gi 55822365 1 MNISEIKDLLAQFDASTLREFSYKNNGEELNLSK.[19].SSPQ.[4].PVAAPAA.[1].ETPA.[1].PVEEA 79 gi 116627300 1 MNISEIKDLLAQFDASTLREFSYKNNGEELNLSK.[19].SSPQ.[4].PVAAPAA.[1].ETPA.[1].PVEEA 79
gi 261417741 87 ET.[11].GNLHQITSPMVGTFYAAPAPDKPPYVKPGDKVKKDTVVCIIEAMKLFNEIEAEVDGEIVEVLVQNGQL 167 gi 146319455 76 AS.[ 2].AEGAVVESPLVGVAYLSPAPDKPAFVAVGDTVKKGQTLMIIEAMKVMNEVPADRDGVVTEILVANQML 147 gi 146321652 76 AS.[ 2].AEGAVVESPLVGVAYLSPAPDKPAFVAVGDTVKKGQTLMIIEAMKVMNEVPADRDGVVTEILVANQDV 147 gi 253752470 76 AS.[ 2].AEGAVVESPLVGVAYLSPAPDKPAFVAVGDTVKKGQTLMIIEAMKVMNEVPADRDGVVTEILVANQDV 147 gi 253754296 76 AS.[ 2].AEGAVVESPLVGVAYLSPAPDKPAFVAVGDTVKKGQTLMIIEAMKVMNEVPADRDGVVTEILVANQDV 147 gi 253756230 76 AS.[ 2].AEGAVVESPLVGVAYLSPAPDKPAFVAVGDTVKKGQTLMIIEAMKVMNEVPADRDGVVTEILVANQDV 147 gi 288904558 75 ET.[ 7].AEGDEVTSPLVGVAYLAPAPDKPAFVSVGDSVKKGQTLLIIEAMKVMNEIPAPNDGIVTEIMVNNEDV 151 gi 55820480 80 SA.[ 2].AEGEVVESPLVGVAYLSPSPEKPAFVSVGDTVKKGQTLLIVEAMKVMNEVPAPKDGVITEILVANEEV 151 gi 55822365 80 SA.[ 2].AEGEVVESPLVGVAYLSPSPEKPAFVSVGDTVKKGQTLLIVEAMKVMNEVPAPKDGVITEILVANEEV 151 gi 116627300 80 SA.[ 2].AEGEVVESPLVGVAYLSPSPEKPAFVSVGDTVKKGQTLLIVEAMKVMNEVPAPKDGVITEILVANEEV 151
gi 261417741 168 .[11]. 178 gi 146319455 gi 146321652 148 .[11]. 158 gi 253752470 148 .[11]. 158 gi 253754296 148 .[11]. 158 gi 253756230 148 .[11]. 158 gi 288904558 152 .[11]. 162 gi 55820480 152 .[11]. 162 gi 55822365 152 .[11]. 162 gi 116627300 152 .[11]. 162
*GC% Content for gene: 6.45%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
ATGGATTTTA AAGGCATTGA AAATTTGATC AAAGCGATGT CTGAGAGCAA CCTGTCCTCA ATGGATATTG AATACAATGG TATTGCGATT AAAATGAAAA AGGAAAACAA CAAAATTTAT AAACAAGAAA CTATCTCACA AGAGTATGAG AAAGAAAATC GGGATAATAT CGTAGAGGAA AAGAAACTGG ACCTGTTGTC AAACGAAGAG AAAACGGGTA TCGCGATTGC CGATAATCTT ATTGAGATTG TTAGCCCAAT CGTTGGTACG TTCTATGAGT CCCCGGGCGT GGACAAAAAA CCATATGCGA AAGCGGGCGA CAAAGTGAAA AAAGGGGATA CCGTGTGCAT CGTTGAAGCT ATGAAAGTAA TGAACGAGAT TGAAGCGGAA GTGGATGGGG AAATTGTCGA AGTCCTGGTG GAGAATGAAC AAATGGTTCA ATACGGTGAA GTTCTGTTTA AGATCAAACC CCTG
*GC% Content for gene (codon optimized): 39.87%

Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.

Primer design results for 'tail' primers (this is just 2 sequences):
**